Computerised cognitive training for 12 or more weeks for maintaining cognitive function in cognitively healthy people in late life.
Increasing age is associated with a natural decline in cognitive function and is the greatest risk factor for dementia. Cognitive decline and dementia are significant threats to independence and quality of life in older adults. Therefore, identifying interventions that help to maintain cognitive function in older adults or that reduce the risk of dementia is a research priority. Cognitive training uses repeated practice on standardised exercises targeting one or more cognitive domains and may be intended to improve or maintain optimal cognitive function. This review examines the effects of computerised cognitive training interventions lasting at least 12 weeks on the cognitive function of healthy adults aged 65 or older and has formed part of a wider project about modifying lifestyle to maintain cognitive function. We chose a minimum 12 weeks duration as a trade-off between adequate exposure to a sustainable intervention and feasibility in a trial setting. To evaluate the effects of computerised cognitive training interventions lasting at least 12 weeks on cognitive function in cognitively healthy people in late life. We searched to 31 March 2018 in ALOIS (www.medicine.ox.ac.uk/alois), and we performed additional searches of MEDLINE, Embase, PsycINFO, CINAHL, ClinicalTrials.gov, and the WHO Portal/ICTRP (www.apps.who.int/trialsearch), to ensure that the search was as comprehensive and as up-to-date as possible to identify published, unpublished, and ongoing trials. We included randomised controlled trials (RCTs) and quasi-RCTs, published or unpublished, reported in any language. Participants were cognitively healthy people, and at least 80% of the study population had to be aged 65 or older. Experimental interventions adhered to the following criteria: intervention was any form of interactive computerised cognitive intervention - including computer exercises, computer games, mobile devices, gaming console, and virtual reality - that involved repeated practice on standardised exercises of specified cognitive domain(s) for the purpose of enhancing cognitive function; the duration of the intervention was at least 12 weeks; cognitive outcomes were measured; and cognitive training interventions were compared with active or inactive control interventions. We performed preliminary screening of search results using a 'crowdsourcing' method to identify RCTs. At least two review authors working independently screened the remaining citations against inclusion criteria. At least two review authors also independently extracted data and assessed the risk of bias of included RCTs. Where appropriate, we synthesised data in random-effects meta-analyses, comparing computerised cognitive training (CCT) separately with active and inactive controls. We expressed treatment effects as standardised mean differences (SMDs) with 95% confidence intervals (CIs). We used GRADE methods to describe the overall quality of the evidence for each outcome. We identified eight RCTs with a total of 1183 participants. The duration of the interventions ranged from 12 to 26 weeks; in five trials, the duration of intervention was 12 or 13 weeks. The included studies had moderate risk of bias, and the overall quality of evidence was low or very low for all outcomes. We compared CCT first against active control interventions, such as watching educational videos. Negative SMDs favour CCT over control. Trial results suggest slight improvement in global cognitive function at the end of the intervention period (12 weeks) (standardised mean difference (SMD) -0.31, 95% confidence interval (CI) -0.57 to -0.05; 232 participants; 2 studies; low-quality evidence). One of these trials also assessed global cognitive function 12 months after the end of the intervention; this trial provided no clear evidence of a persistent effect (SMD -0.21, 95% CI -0.66 to 0.24; 77 participants; 1 study; low-quality evidence). CCT may result in little or no difference at the end of the intervention period in episodic memory (12 to 17 weeks) (SMD 0.06, 95% CI -0.14 to 0.26; 439 participants; 4 studies; low-quality evidence) or working memory (12 to 16 weeks) (SMD -0.17, 95% CI -0.36 to 0.02; 392 participants; 3 studies; low-quality evidence). Because of the very low quality of the evidence, we are very uncertain about the effects of CCT on speed of processing and executive function. We also compared CCT to inactive control (no interventions). We found no data on our primary outcome of global cognitive function. At the end of the intervention, CCT may lead to slight improvement in episodic memory (6 months) (mean difference (MD) in Rivermead Behavioural Memory Test (RBMT) -0.90 points, 95% confidence interval (CI) -1.73 to -0.07; 150 participants; 1 study; low-quality evidence) but can have little or no effect on executive function (12 weeks to 6 months) (SMD -0.08, 95% CI -0.31 to 0.15; 292 participants; 2 studies; low-quality evidence), working memory (16 weeks) (MD -0.08, 95% CI -0.43 to 0.27; 60 participants; 1 study; low-quality evidence), or verbal fluency (6 months) (MD -0.11, 95% CI -1.58 to 1.36; 150 participants; 1 study; low-quality evidence). We could not determine any effects on speed of processing because the evidence was of very low quality. We found no evidence on quality of life, activities of daily living, or adverse effects in either comparison. We found low-quality evidence suggesting that immediately after completion of the intervention, small benefits of CCT may be seen for global cognitive function when compared with active controls, and for episodic memory when compared with an inactive control. These benefits are of uncertain clinical importance. We found no evidence that the effect on global cognitive function persisted 12 months later. Our confidence in the results was low, reflecting the overall quality of the evidence. In five of the eight trials, the duration of the intervention was just three months. The possibility that more extensive training could yield larger benefit remains to be more fully explored. We found substantial literature on cognitive training, and collating all available scientific information posed problems. Duration of treatment may not be the best way to categorise interventions for inclusion. As the primary interest of older people and of guideline writers and policymakers involves sustained cognitive benefit, an alternative would be to categorise by length of follow-up after selecting studies that assess longer-term effects.


Expression of progesterone receptor A and B isoforms in low-grade endometrial stromal sarcoma.
The progesterone receptor (PR) exists as two isoforms, PRA and PRB. In vitro studies have shown that these proteins are functionally distinct, suggesting that their relative expression can influence progesterone response. Low-grade endometrial stromal sarcoma (LGESS) is an uncommon tumor that usually expresses PR. In normal endometrial stroma, both PR isoforms are present with PRA predominant throughout the menstrual cycle. The relative expression of PRA and PRB in LGESS has not been previously reported. All nine cases of primary LGESS (seven uterine, two extrauterine) expressed PRB. Eight tumors also contained PRA and it was the predominant isoform in seven cases. These tumors had similar histopathologic appearances, whereas a case with approximately equal PR isoform expression showed features of sex cord or smooth muscle differentiation. An extrauterine tumor expressing only PRB had myxoid stroma. Recurrent tumor in two cases, which expressed predominantly PRA in the primary, contained reduced levels of PR consisting predominantly or entirely of PRB after prolonged interval progestin therapy. Most primary LGESSs showed PR isoform expression similar to normal endometrial stroma, consistent with the highly differentiated phenotype of this tumor. Variant differentiation or disease recurrence was accompanied by an altered PR isoform profile that could impact on hormone response.


Variability in surface infrared reflectance of thirteen nitrile rubber gloves at key wavelengths for analysis of captan.
The aim of this study was to investigate the surface variability of 13 powder-free, unlined, and unsupported nitrile rubber gloves using attenuated total reflection Fourier transform infrared (ATR-FT-IR) spectrophotometry at key wavelengths for analysis of captan contamination. The within-glove, within-lot, and between-lot variability was measured at 740, 1124, 1252, and 1735 cm(-1), the characteristic captan reflectance minima wavelengths. Three glove brands were assessed after conditioning overnight at relative humidity (RH) values ranging from 2 +/- 1 to 87 +/- 4% and temperatures ranging from -8.6 +/- 0.7 to 59.2 +/- 0.9 degrees C. For all gloves, 1735 cm(-1) provided the lowest background absorbance and greatest potential sensitivity for captan analysis on the outer glove surface: absorbances ranged from 0.0074 +/- 0.0005 (Microflex) to 0.0195 +/- 0.0024 (SafeSkin); average within-glove coefficients of variation (CV) ranged from 2.7% (Best, range 0.9-5.3%) to 10% (SafeSkin, 1.2-17%); within-glove CVs greater than 10% were for one brand (SafeSkin); within-lot CVs ranged from 2.8% (Best N-Dex) to 28% (SafeSkin Blue); and between-lot variation was statistically significant (p < or = 0.05) for all but two SafeSkin lots. The RH had variable effects dependent on wavelength, being minimal at 1735, 1252, and 1124 cm(-1) and highest at 3430 cm(-1) (O-H stretch region). There was no significant effect of temperature conditioning. Substantial within-glove, within-lot, and between-lot variability was observed. Thus, surface analysis using ATR-FT-IR must treat glove brands and lots as different. ATR-FT-IR proved to be a useful real-time analytical tool for measuring glove variability, detecting surface humidity effects, and choosing selective and sensitive wavelengths for analysis of nonvolatile surface contaminants.


Case Report: An Atypical Erysipeloid Presentation of Cutaneous Leishmaniasis from the Hilly Region of Nepal.
Cutaneous leishmaniasis, which is the most common form of leishmaniasis, classically presents as small erythematous papules and nodules that develop into ulcers with indurated, raised outer borders. However, lesions of cutaneous leishmaniasis can have pleomorphic and atypical presentations. The erysipeloid form is one of the rare, atypical presentations of cutaneous leishmaniasis. Reported is a case of a 58-year-old man from the hilly region of Nepal who presented with an atypical erythematous and edematous plaque over the left antecubital fossa. Cutaneous leishmaniasis was not considered as an initial diagnosis because of the atypical appearance of the lesion as well as his residence in the hilly region of Nepal. The diagnosis was made after detection of amastigotes on histopathological examination of a cutaneous biopsy specimen. There was complete regression of the lesion after treatment with oral miltefosine followed by oral fluconazole. Clinicians should be aware of atypical presentations of cutaneous leishmaniasis and it should be considered in the differential diagnosis, regardless of the presentation or geographic location.


Relative bioactivity of histamine and slow reacting substance of anaphylaxis released during anaphylactic reaction from different tissues of guinea pigs and rats.
Relative amount of histamine and SRS-A released during Schultz Dale reaction were indirectly estimated. Percent block produced by specific antagonists, mepyramine and FPL 55712, on antigen induced response in various tissues was compared with controls. The relative bioactivity of histamine and SRS-A released was 93% and 7%, respectively, in rat lungs: 37% and 63% in guinea pig lungs; 82% and 18% in guinea pig intestine, 59% and 41% in rat intestine and 100% and 0% in skin of guinea pig and rats. Expectedly, individual experiments showed gross variations because anaphylaxis is rarely dose related.


Inhibition of the malate-aspartate shuttle in mouse pancreatic islets abolishes glucagon secretion without affecting insulin secretion.
Altered secretion of insulin as well as glucagon has been implicated in the pathogenesis of Type 2 diabetes (T2D), but the mechanisms controlling glucagon secretion from α-cells largely remain unresolved. Therefore, we studied the regulation of glucagon secretion from αTC1-6 (αTC1 clone 6) cells and compared it with insulin release from INS-1 832/13 cells. We found that INS-1 832/13 and αTC1-6 cells respectively secreted insulin and glucagon concentration-dependently in response to glucose. In contrast, tight coupling of glycolytic and mitochondrial metabolism was observed only in INS-1 832/13 cells. Although glycolytic metabolism was similar in the two cell lines, TCA (tricarboxylic acid) cycle metabolism, respiration and ATP levels were less glucose-responsive in αTC1-6 cells. Inhibition of the malate-aspartate shuttle, using phenyl succinate (PhS), abolished glucose-provoked ATP production and hormone secretion from αTC1-6 but not INS-1 832/13 cells. Blocking the malate-aspartate shuttle increased levels of glycerol 3-phosphate only in INS-1 832/13 cells. Accordingly, relative expression of constituents in the glycerol phosphate shuttle compared with malate-aspartate shuttle was lower in αTC1-6 cells. Our data suggest that the glycerol phosphate shuttle augments the malate-aspartate shuttle in INS-1 832/13 but not αTC1-6 cells. These results were confirmed in mouse islets, where PhS abrogated secretion of glucagon but not insulin. Furthermore, expression of the rate-limiting enzyme of the glycerol phosphate shuttle was higher in sorted primary β- than in α-cells. Thus, suppressed glycerol phosphate shuttle activity in the α-cell may prevent a high rate of glycolysis and consequently glucagon secretion in response to glucose. Accordingly, pyruvate- and lactate-elicited glucagon secretion remains unaffected since their signalling is independent of mitochondrial shuttles.


[Surveillance of nalidixic acid-resistant and extended-spectrum beta-lactamase-producing Salmonella spp. isolated from human feces].
Nalidixic acid (NA)-resistant and extended-spectrum beta-lactamase (ESBL)-producing Salmonella sp. isolates from human specimens are associated with clinical failure or delayed response in subjects treated with fluoroquinolone or third-generation cephalosporins. We studied drug susceptibility in 604 Salmonella enterica isolates from human feces in 2007. Of these, 39 (6.5%) were resistat to NA. Of these, 46% were resistant to two or more drugs and 2% susceptible to NA were resistant to multiple drugs (p < 0.001). Three ESBL-producing Salmonella sp. isolated were of the CTX-M family gene type. One strain of plasmid-mediated AmpC beta-lactamase belonged to the CMY-2 family gene type. Our results thus showed that NA-resistant isolates were resistant to antimicrobial agents and confirmed the presence of a small number of isolates producing ESBL and AmpC beta-lactamase.


Suppression of CX43 expression by miR-20a in the progression of human prostate cancer.
The aberrant expression of microRNAs (miRNAs) has been found in various types of cancer. The present study found miR-20a to be significantly upregulated in prostate cancer compared with normal prostate tissues. The proliferation and colony formation assays revealed that the downregulation of miR-20a by miR-20a inhibitor suppresses the proliferation of MDA-PCa-2b cells in vitro and also inhibits tumor growth in vivo. Furthermore, a gap junction protein, α 1 (CX43), was identified as a direct target gene of miR-20a. The upregulation of CX43 was detected in MDA-PCa-2b cells after treatment with miR-20a inhibitor both in vitro and in vivo. In conclusion, the findings show that miR-20a significantly contributes to the progression of prostate cancer by targeting CX43.


Rare syndromes of the head and face-Pierre Robin sequence.
Pierre Robin sequence (PRS) is an association of clinical features consisting of mandibular hypoplasia, cleft secondary palate, and glossoptosis leading to obstructive apnea and feeding difficulties. PRS can occur as an isolated condition or can be found in association with a range of other features in a number of conditions including Treacher collins and Stickler syndromes. The frequent association of the PRS triad suggests a common underlying developmental mechanism which impacts on each of these tissues. Isolated PRS is typically sporadic but when familial usually exhibits autosomal dominant inheritance. The term PRS is applied on the basis of the pattern of malformation rather than etiology and growing evidence indicates that the initiating genetic lesion is variable. Various chromosomal anomalies have been associated with PRS including loci on chromosomes 2, 4, and 17. Associations with genes including SOX9, a number of collagen genes and work with animal models suggest the phenotype derives from a cartilage defect during early facial growth. However, alternative theories have been proposed and these highlight the difficulty of characterising congenital anomalies of craniofacial development in which multiple etiologies can result in very similar phenotypes.


Glycopolymer micelles with reducible ionic cores for hepatocytes-targeting delivery of DOX.
A novel galactose-decorated cross-linked micelles (cl-micelles) with ionic cores using cystamine (Cys) as a biodegradable cross-linker was prepared by using block ionomer complexes of poly(ethylene glycol)-b-poly(2-acryloxyethyl-galactose)-b-poly(acrylic acid) (PEG-b-PAEG-b-PAA) and Ca(2+) (PEG-b-PAEG-b-PAA cl-micelles/Cys). Doxorubicin (DOX) was successfully incorporated into the ionic cores of such micelles via electrostatic interactions. Proton nuclear magnetic resonance spectrum and Fourier transform infrared spectrometer indicated galactose ligands were exposed at the micellar surface. The micelles were spherical in shape, with an average size of 100nm. The in vitro release studies confirmed that DOX-loaded PEG-b-PAEG-b-PAA cl-micelles/Cys accomplished rapid drug release under reducing condition. Remarkably, PEG-b-PAEG-b-PAA cl-micelles/Cys efficiently delivered and released DOX into the cell nucleus of HepG2 cells, and the intensity of fluorescence observed in HepG2 cells was stronger than that incubated with the micelles without galactose ligands. In contrast, little fluorescence was observed in NIH3T3 cells after incubation with PEG-b-PAEG-b-PAA cl-micelles/Cys. Interestingly, cytotoxicity assays showed that DOX-loaded PEG-b-PAEG-b-PAA cl-micelles/Cys retained higher cell inhibition efficiency in HepG2 cells as compared with NIH3T3 cells, and were more potent than the micelles without galactose ligands and the micelles with non degradable cross-links. These results indicate that PEG-b-PAEG-b-PAA cl-micelles/Cys have great potential in liver tumor-targeted chemotherapy.


Unprecedented redox-driven ligand ejection in nickel(II)-diiminosemiquinonate radical complexes.
Nickel(II) complexes of (M : L) stoichiometries 1 : 1 (1) and 1 : 2 (2) were prepared from a polydentate ligand involving diiminosemiquinonate radicals. Both were characterized by X-ray diffraction, Vis-NIR and EPR spectroscopy as well as electrochemistry. Ligand-centered oxidation of 1 promotes ligand ejection to give 2.


Adolescents Exiting Homelessness Over Two Years: The Risk Amplification and Abatement Model.
The Risk Amplification and Abatement Model (RAAM), demonstrates that negative contact with socializing agents amplify risk, while positive contact abates risk for homeless adolescents. To test this model, the likelihood of exiting homelessness and returning to familial housing at 2 years and stably exiting over time are examined with longitudinal data collected from 183 newly homeless adolescents followed over 2 years in Los Angeles, CA. In support of RAAM, unadjusted odds of exiting at 2 years and stably exiting over2 years revealed that engagement with pro-social peers, maternal social support, and continued school attendance all promoted exiting behaviors. Simultaneously, exposure to family violence and reliance on shelter services discouraged stably exiting behaviors. Implications for family-based interventions are proposed.


RNAase-III enzyme Dicer maintains signaling pathways for differentiation and survival in mouse cortical neural stem cells.
An important function of the RNAase-III enzyme Dicer is to process microRNA precursors into ~22-nucleotide non-coding small RNAs. But little is known about the role of Dicer in mammalian brain formation and neural stem cell (NSC) development. Here we show that Dicer plays a crucial role in controlling mouse cortical NSC development. We found that Dicer function is essential for expanding cortical neural progenitors and NSCs. We have identified a population of Dicer-deficient NSCs that can self-renew, and that display normal karyotype and heterochromatin protein expression levels but show enlarged nuclei. Dicer-deficient NSCs display abnormal differentiation and undergo cell death when mitogens are withdrawn. Dicer deletion affects the levels of many proteins, as revealed by a mass spectrometry proteomic approach. We have found that an increase of anti-survival and/or pro-apoptosis proteins and a decrease of pro-survival and/or anti-apoptosis proteins contribute to the cell death of Dicer-deficient NSCs, implying a general role for Dicer in protecting cells from apoptosis. Our results demonstrate important functions for Dicer in regulating NSC development by maintaining proper signaling pathways related to cell survival and differentiation.


Effect of Rothia mucilaginosa enzymes on gliadin (gluten) structure, deamidation, and immunogenic epitopes relevant to celiac disease.
Rothia mucilaginosa, a natural microbial inhabitant of the oral cavity, cleaves gluten (gliadin) proteins at regions that are resistant to degradation by mammalian enzymes. The aim of this study was to investigate to what extent the R. mucilaginosa cell-associated enzymes abolish gliadin immunogenic properties. Degradation of total gliadins and highly immunogenic gliadin 33-mer or 26-mer peptides was monitored by SDS-PAGE and RP-HPLC, and fragments were sequenced by liquid chromatography and electrospray ionization tandem mass spectrometer (LC-ESI-MS/MS). Peptide deamidation by tissue transglutaminase (TG2), a critical step in rendering the fragments more immunogenic, was assessed by TG2-mediated cross-linking to monodansyl cadaverine (MDC), and by a +1-Da mass difference by LC-ESI-MS. Survival of potential immunogenic gliadin epitopes was determined by use of the R5 antibody-based ELISA. R. mucilaginosa-associated enzymes cleaved gliadins, 33-mer and 26-mer peptides into smaller fragments. TG2-mediated cross-linking showed a perfect inverse relationship with intact 33-mer and 26-mer peptide levels, and major degradation fragments showed a slow rate of MDC cross-linking of 6.18 ± 2.20 AU/min compared with 97.75 ± 10.72 and 84.17 ± 3.25 AU/min for the intact 33-mer and 26-mer, respectively, which was confirmed by reduced TG2-mediated deamidation of the fragments in mass spectrometry. Incubation of gliadins with Rothia cells reduced R5 antibody binding by 20, 82, and 97% after 30 min, 2 h, and 5 h, respectively, which was paralleled by reduced reactivity of enzyme-treated 33-mer and 26-mer peptides in the R5 competitive ELISA. Our broad complementary approach to validate gluten degrading activities qualifies R. mucilaginosa-associated enzymes as promising tools to neutralize T cell immunogenic properties for treatment of celiac disease.


First steps to understand productivity loss in Portuguese patients with rheumatoid arthritis.
Rheumatoid Arthritis (RA), in consequence of joint inflammation and damage, is known to lead to productivity loss. Developments during the 21st century, such as biologic treatments, allowed remission/low disease activity to be achieved in more RA patients. Despite this, evidence is equivocal concerning work outcomes improvement. Our goal was to evaluate work status and productivity in a Portuguese population with RA from 3 Rheumatology Departments through the questionnaire Work Productivity and Activity Impairment General Health (WPAI).


[Effect of anorexia and neuropathic pain induced by cisplatin on hindlimb muscles of rat].
The purpose of this study was to examine the effect of anorexia and neuropathic pain induced by cisplatin on hindlimb muscles of rats. Adult male Sprague-Dawley rats were divided into two groups, a cisplatin-treated group (n=10) and a control group (n=10). In the cisplatin-treated group, cisplatin at a dose of 2 mg/kg was injected intraperitoneally two times a week up to a cumulative dose of 20 mg/kg over 5 weeks, and in the control group saline (0.9% NaCl) was injected intraperitoneally at the same dose and duration as the cisplatin-treated group. At 34 days all rats were anesthetized, after which the soleus and plantaris muscles were dissected. Withdrawal threshold, body weight, food intake, activity, muscle weight, Type I and II fiber cross-sectional areas and myofibrillar protein content of the dissected muscles were determined. Compared with the control group, the cisplatin-treated group showed significant decreases (p<.05) in withdrawal threshold, activity, food intake, body weight, Type I and II fiber cross-sectional areas, myofibrillar protein content and weight of the soleus and plantaris muscles. Muscular atrophy in hindlimb occurs due to anorexia and neuropathic pain induced by the cisplatin treatment.


Reduced receptor binding by a human interferon-gamma fragment lacking 11 carboxyl-terminal amino acids.
Treatment of recombinant human interferon-gamma (IFN-gamma) with either 1) the arginine-specific proteases clostripain or submaxillaris protease or 2) the broadly specific enzyme pronase produced a stable fragment with m.w. of 15,500. Structural analysis showed that the cleavage occurred between residues 129 and 130 and thus produced a fragment lacking only 11 carboxyl-terminal amino acids. The fragmented and untreated molecules showed identical amino-terminal amino acid sequences and were equally reactive with either polyclonal or monoclonal anti-IFN-gamma. IFN-gamma lacking carboxyl-terminal amino acids displayed a 1000- to 2000-fold reduction in its capacity to bind to cellular IFN-gamma receptors at 4 degrees C. Functionally the fragment showed a 50-fold reduction in its ability to induce antiviral activity in fibroblasts and a 10-fold reduction in its ability to induce Fc receptors on the human histiocytic lymphoma cell line U937. These results thus suggest that the carboxyl terminus of human IFN-gamma contributes significantly to the formation of the receptor-binding site of the molecule.


[Medical aspects on sauna bathing].
The literature describing medical aspects on sauna bathing is briefly reviewed. The circulatory effects of heating and subsequent cooling related to sauna bath differ with age, underlying medical condition and use of drugs. The circulatory changes may often be beneficial but can sometimes prove dangerous. More than 2000 sauna bathers were followed for around 20 years in a Finnish population study. The investigators have recently reported health benefits of abundant sauna use for individuals concerning hypertension, ischemic heart disease, dementia and certain pulmonary diseases. Others have recommended the use of sauna for patients with heart failure. Fatal events in sauna are very uncommon and often linked to the use of alcohol, to ischemic heart disease and to bathing alone. Medical conditions in which sauna bathing should be avoided are shortly reviewed. Further studies are urgently needed for illustration of the possible benefit of sauna bath at different medical conditions.


Epistasis between mutations is host-dependent for an RNA virus.
How, and to what extent, does the environment influence the way mutations interact? Do environmental changes affect both the sign and the magnitude of epistasis? Are there any correlations between environments in the variability, sign or magnitude of epistasis? Very few studies have tackled these questions. Here, we addressed them in the context of viral emergence. Most emerging viruses are RNA viruses with small genomes, overlapping reading frames and multifunctional proteins for which epistasis is abundant. Understanding the effect of host species in the sign and magnitude of epistasis will provide insights into the evolutionary ecology of infectious diseases and the predictability of viral emergence.


Relaxin activates the L-arginine-nitric oxide pathway in vascular smooth muscle cells in culture.
The peptide hormone relaxin (RLX) has been shown to elicit a powerful vasodilatory response in several target organs. This response is mediated by the stimulation of intrinsic nitric oxide (NO) generation. The present study was designed to clarify whether RLX directly promotes the relaxation of vascular smooth muscle cells through stimulation of NO generation. Vascular smooth muscle cells from bovine aortas were incubated with RLX at concentrations ranging from 1 nmol/L to 1 micromol/L. The expression and activity of NO synthase, production of NO, and the intracellular levels of cGMP and Ca2+ were determined. The cell morphology and signal transduction mechanisms of these bovine aortic smooth muscle cells in response to RLX were also studied. RLX stimulated the expression of immunoreactive inducible NO synthase and increased significantly and in a concentration-related fashion inducible NO synthase activity, NO generation, and intracellular cGMP levels. Concurrently, RLX significantly decreased cytosolic Ca2+ concentrations and caused changes in cell shape and the actin cytoskeleton that were consistent with cell relaxation. The signal transduction mechanisms leading to the enhanced expression of inducible NO synthase protein and activity caused by RLX involve the activation of tyrosine kinase, phosphatidylcholine-phospholipase C, and the transcription factor nuclear factor-kappaB, similar to bacterial endotoxins and proinflammatory cytokines. This study suggests that RLX is an endogenous agent capable of regulating vascular tone by activation of the L-arginine-NO pathway in vascular smooth muscle cells.


Iatrogenic FDG foci in the lungs: a pitfall of PET image interpretation.
2-[F-18]-fluoro-2-deoxy-D-glucose (FDG) positron emission tomography (PET) has become an important staging modality for many tumors, including bronchial carcinoma; however it is important to know that there are several pitfalls in PET image interpretation. In this report we demonstrate three cases in which focal intrapulmonary FDG uptake could possibly represent iatrogenic microembolism. These FDG accumulations would have been interpreted as malignant tumor mass in the lung if no anatomic correlation would have been performed. For this reason, we further present an integrated PET/CT scanner, which recently has been introduced. This correlation of molecular and morphological information enables the specification of the FDG-PET findings.


A proteolytic fragment of Mcl-1 exhibits nuclear localization and regulates cell growth by interaction with Cdk1.
Mcl-1 (myeloid cell leukaemia-1) is a Bcl-2 family member with short-term pro-survival functions but whose other functions, demonstrated by embryonic lethality of knockout mice, do not involve apoptosis. In the present study, we show a cell-cycle-regulatory role of Mcl-1 involving a shortened form of the Mcl-1 polypeptide, primarily localized to the nucleus, which we call snMcl-1. snMcl-1 interacts with the cell-cycle-regulatory protein Cdk1 (cyclin-dependent kinase 1; also known as cdc2) in the nucleus, and Cdk1 bound to snMcl-1 was found to have a lower kinase activity. The interaction with Cdk1 occurs in the absence of its cyclin partners and is enhanced on treatment of cells with G2/M blocking agents, but not by G1/S blocking. The snMcl-1 polypeptide is present during S and G2 phases and is negligible in G1. Overexpression of human Mcl-1 in a murine myeloid progenitor cell line resulted in a lower rate of proliferation. Furthermore, Mcl-1-overexpressing cells had lower total Cdk1 kinase activity compared with parental cells, in both anti-Cdk1 and anti-cyclin B1 immunoprecipitates. The latter results suggest that binding to snMcl-1 alters the ability of Cdk1 to bind its conventional partner, cyclin B1. Given the important role of Cdk1 in progression through G2 and M phases, it is probable that the inhibition of Cdk1 activity accounts for the inhibitory effect of Mcl-1 on cell growth.


Observations on the classification of the inflammatory myopathies.
This brief review considers historical approaches to the classification of the inflammatory myopathies. The last 25 years have seen advances in our knowledge of the underlying immune mechanism but the initial trigger for the idiopathic inflammatory myopathies remains unknown. Existing classifications have their limitations, but with the absence of a "gold standard" a definitive classification is not yet possible. Despite these problems, a working classification is possible that is valuable for everyday clinical practice.


Energy dependence of lung phosphatidylcholine biosynthesis studied with CO hypoxia.
The energy dependence of lung phosphatidylcholine (PC) biosynthesis was evaluated by measuring incorporation of radiolabeled precursor substrates by the isolated perfused lung during carbon monoxide (CO)-induced hypoxia. Ventilation with gases containing 75 or 90% CO (plus 5% O2 and 5% CO2) resulted in a mean decrease of lung ATP content by 29 and 53%, respectively, and a significant decrease of [U-14C]glucose incorporation into free fatty acids and the fatty acyl moiety of lung PC. [9,10-3H]palmitate incorporation into lung PC was also decreased by hypoxia but [methyl-14C] choline incorporation was unaffected. For disaturated PC, incorporation of all three substrates was significantly depressed during CO ventilation. These results indicate that lung synthesis of PC is inhibited by alterations of lung energy state. The reactions in lung PC metabolism that are inhibited by this degree of energy limitation are the de novo synthesis of fatty acids from glucose, the acylation of exogenous palmitate, and the remodeling of PC into disaturated PC by the deacylation-reacylation pathway.


Selective aortic arch perfusion: a first-in-human observational cadaveric study.
Selective aortic arch perfusion (SAAP) is a novel endovascular technique that combines thoracic aortic occlusion with extracorporeal perfusion of the brain and heart. SAAP may have a role in both haemorrhagic shock and in cardiac arrest due to coronary ischaemia. Despite promising animal studies, no data is available that describes SAAP in humans. The primary aim of this study was to assess the feasibility of selective aortic arch perfusion in humans. The secondary aim of the study was to assess the feasibility of achieving direct coronary artery access via the SAAP catheter as a potential conduit for salvage percutaneous coronary intervention. Using perfused human cadavers, a prototype SAAP catheter was inserted into the descending aorta under fluoroscopic guidance via a standard femoral percutaneous access device. The catheter balloon was inflated and the aortic arch perfused with radio-opaque contrast. The coronary arteries were cannulated through the SAAP catheter. The procedure was conducted four times. During the first two trials the SAAP catheter was passed rapidly and without incident to the intended descending aortic landing zone and aortic arch perfusion was successfully delivered via the device. The SAAP catheter balloon failed on the third trial. On the fourth trial the left coronary system was cannulated using a 5Fr coronary guiding catheter through the central SAAP catheter lumen. For the first time using a perfused cadaveric model we have demonstrated that a SAAP catheter can be easily and safely inserted and SAAP can be achieved using conventional endovascular techniques. The SAAP catheter allowed successful access to the proximal aorta and permitted retrograde perfusion of the coronary and cerebral circulation.


Effects of CRH and LHRH on rat septo-hippocampal neurons.
Electrical activities of septum-diagonal band neurons (SD neurons) antidromically activated by electrical stimulation of the fimbria were recorded in urethane anesthetized male rats. The mean basal firing rate of antidromically identified SD neurons (2.3 Hz) was not different from that of unidentified ones (2.2 Hz). The mean latency of antidromic activation by the fimbria stimulation was 4.2 ms. Iontophoretically applied CRH and LHRH affected 60% and 40% of identified SD neurons, but only 19% and 13% of unidentified SD neurons, respectively. The ratio of response to no response to both peptides was significantly different in unidentified SD neurons from that in identified SD neurons. These results suggest that SD neurons mediate the effect of CRH and LHRH on hippocampal neurons.


Estrogen acutely abolishes abnormal cold-induced coronary constriction in men.
Ambient cold exposure may induce myocardial ischemia by precipitating coronary artery constriction and a decrease in coronary blood flow. Estrogen has vasoactive properties that may prevent abnormal coronary constriction in a sex-independent manner. The purpose of this study is to determine whether estrogen acutely abolishes abnormal coronary responses to cold exposure in men. Randomized, double-blinded placebo-controlled clinical trial. Cardiac catheterization laboratory. Men referred for routine diagnostic coronary angiography who exhibit abnormal coronary artery constriction in response to a 90-s cold pressor test (CPT). Intravenous conjugated estrogens (1.25 mg) vs. placebo. Rate-pressure product, coronary cross-sectional area (CSA), and coronary blood flow responses to the CPT were measured before and 15 min after intervention. In 12 men with CPT-induced coronary constriction who were assigned to estrogen, CPT induced a mean 21.8% decrease in coronary CSA (p < 0.01) and a nonsignificant change in coronary flow. After estrogen, the repeated CPT induced a 16.3% increase in CSA (p < 0.01) and a 54.9% increase in flow (p < 0.01). CSA and coronary flow responses to CPT were significantly different before and after estrogen (p < 0.01). In contrast, placebo was not associated with changes in CSA or coronary flow responses to CPT in eight men. In men, conjugated estrogens acutely abolish abnormal coronary constriction and improve coronary blood flow responses to an exogenous cold stimulus. These results suggest that estrogen favorably alters coronary vasoreactivity in men.


Recurrent genomic rearrangements in primary testicular lymphoma.
Primary testicular diffuse large B cell lymphoma (PTL) is an aggressive malignancy that occurs in the immune-privileged anatomical site of the testis. We have previously shown that structural genomic rearrangements involving the MHC class II transactivator CIITA and programmed death ligands (PDLs) 1 and 2 are frequent across multiple B cell lymphoma entities. Specifically in PTL, we found rearrangements in the PDL locus by fluorescence in situ hybridization (FISH). However, breakpoint anatomy and rearrangement partners were undetermined, while CIITA rearrangements had not been reported previously in PTL. Here, we performed bacterial artificial chromosome capture sequencing on three archival, formalin-fixed, paraffin-embedded tissue biopsies, interrogating 20 known rearrangement hotspots in B cell lymphomas. We report novel CIITA, FOXP1 and PDL rearrangements involving IGHG4, FLJ45248, RFX3, SMARCA2 and SNX29. Moreover, we present immunohistochemistry data supporting the association between PDL rearrangements and increased protein expression. Finally, using FISH, we show that CIITA (8/82; 10%) and FOXP1 (5/74; 7%) rearrangements are recurrent in PTL. In summary, we describe rearrangement frequencies and novel rearrangement partners of the CIITA, FOXP1 and PDL loci at base-pair resolution in a rare, aggressive lymphoma. Our data suggest immune-checkpoint inhibitor therapy as a promising intervention for PTL patients harbouring PDL rearrangements.


Patients, intimate partners and family experiences of implantable cardioverter defibrillators: qualitative systematic review.
This paper is a report of an interpretive review of qualitative research on how an implantable cardioverter defibrillator affects adult recipients and their significant others. An implantable cardioverter defibrillator detects pathological cardiac rhythms and automatically converts the rhythm with electrical counter shocks. A systematic literature search was conducted for qualitative research papers published between January 1999 and January 2009. PubMed, Medline, ISI Web of Knowledge and CINAHL databases were searched with the following key words: internal defibrillator, implantable defibrillator and qualitative research. Twenty-two papers were included. The critical appraisal skills programme and prompts were used to appraise studies. Thematic analysis and synthesis approaches were used to interpret evidence. People with an implantable cardioverter defibrillator were found to experience physical, psychological and social changes. Shocks produce fear and anxiety, affecting relationships and sexual relations. The use of support groups and the use of the Internet are important in helping adjustment to an implantable cardioverter defibrillator. Women's responses to an implantable cardioverter defibrillator appear different than men's responses and include concerns about physical appearance and relationship issues. Postdischarge follow-up and educational programmes are still underdeveloped. Patients need additional education, support and follow-up care after hospital discharge. Patients and significant others benefit from collaboration between patient associations and healthcare professional societies. Future research is needed to identify the specific challenges that women recipients face.


[Self-reported diabetes mellitus in the city of São Paulo: prevalence and inequality]
This report analyzes characteristics of self-reported diabetes mellitus in the city of São Paulo, Brazil. The data were obtained from the Brazilian Multicenter Study on Prevalence of Diabetes Mellitus, a household survey performed in 1986-88. The São Paulo sample consisted of 2,007 individuals aged 30-69 years, of both sexes, selected from three areas with distinct socio-economic levels. The estimated prevalence using a 75g glucose load and measurement of two-hour capillary glycemia was 9.7%. Prevalence of self-reported diabetes was 4.7% and increased with age and presence of family history of diabetes. There was a significant difference between sexes (3.5% in men and 5.7% in women), with higher rates of self-reported diabetes at higher economic levels among men and higher rates at lower socio-economic levels among women.


Rational Design and Reticulation of Infinite qbe Rod Secondary Building Units into Metal-Organic Frameworks through a GlobalDesymmetrization Approach for InverseC3H8/C3H6Separation.
The development of reticular chemistry has enabled the construction of a large array of metal-organic frameworks (MOFs) with diverse net topologies and functions. However, dominating this class of materials are those built from discrete/finite secondary building units (SBUs), yet the designed synthesis of frameworks involving infinite rod-shaped SBUs remain underdeveloped. Here, by virtue of a global linker desymmetrization approach, we successfully targeted a novel Cu-MOF (Cu-ASY) incorporating infinite Cu-carboxylate rod SBUs with its structure determined by micro electron diffraction (MicroED) crystallography. Interestingly, the rod SBU can be simplified as a unique cylindric sphere packing qbe tubule made of [43.62] tiles, which further connect the tritopic linkers to give a newly discovered 3,5-connected gfc net. Cu-ASY is a permanent ultramicroporous material featuring 1D channels with highly inert surfaces and shows a preferential adsorption of propane (C3H8) over propene (C3H6). The efficiency of C3H8 selective Cu-ASY is validated by multicycle breakthrough experiments, giving C3H6 productivity of 2.2 L/kg. Density functional theory (DFT) calculations reveal that C3H8 molecules form multiple C-H···π and atypical C-H···H-C van der Waals interactions with the inner nonpolar surfaces. This work therefore highlights the linker desymmetrization as an encouraging and intriguing strategy for achieving unique MOF structures and properties.


Yoga an effective strategy for self-management of stress-related problems and wellbeing during COVID19 lockdown: A cross-sectional study.
This cross-sectional research aims to study the effect of yoga practice on the illness perception, and wellbeing of healthy adults during 4-10 weeks of lockdown due to COVID19 outbreak. A total of 668 adults (64.7% males, M = 28.12 years, SD = 9.09 years) participated in the online survey. The participants were grouped as; yoga practitioners, other spiritual practitioners, and non-practitioners based on their responses to daily practices that they follow. Yoga practitioners were further examined based on the duration of practice as; long-term, mid-term and beginners. Multivariate analysis indicates that yoga practitioners had significantly lower depression, anxiety, & stress (DASS), and higher general wellbeing (SWGB) as well as higher peace of mind (POMS) than the other two groups. The results further revealed that the yoga practitioners significantly differed in the perception of personal control, illness concern and emotional impact of COVID19. However, there was no significant difference found for the measure of resilience (BRS) in this study. Yoga practitioners also significantly differed in the cognitive reappraisal strategy for regulating their emotions than the other two groups. Interestingly, it was found that beginners -those who had started practicing yoga only during the lockdown period reported no significant difference for general wellbeing and peace of mind when compared to the mid- term practitioner. Evidence supports that yoga was found as an effective self- management strategy to cope with stress, anxiety and depression, and maintain wellbeing during COVID19 lockdown.


A sophisticated programmable miniaturised pump for insulin delivery.
We have conceived a truly pre-programmable infusion system usable for intravenous administration of insulin in diabetic subjects. The original system has been built into a small, commercially available, syringe-pump of which only the case and the mechanical parts have been kept. The computing until has a timer, a programmable memory of 512 words by 8 bits and a digital-to-frequency converter to run the motor which drives the syringe. The memory contains 8 profiles of insulin injections stored in digital form over 64 words. Each profile is selected by the patient before eating according to the carbohydrate content of the planned meal and last about two hours, starting from and returning to the basal rate of insulin, at which it remains until next profile selection. Amount, profiles and duration of insulin injection are either mean values deduced from previous studies with a closed-loop artificial pancreas or personally fitted values; they are stored in an instantly replaceable memory cell. This device allows the patient to choose the time, nature and amount of his food intake.


Characterization of human enterovirus71 virus-like particles used for vaccine antigens.
Human enterovirus 71 (EV71) is a major causative pathogen of hand, foot and mouth disease (HFMD) and has caused outbreaks with significant mortality among young children in the Asia-Pacific region in recent years. Towards developing a vaccine for this disease, we have expressed and purified EV71 virus-like particles (VLPs), which resemble the authentic virus in appearance, capsid structure and protein sequence, from insect cells (Sf9) using a multistep chromatography process. We demonstrated intracellular localization of the VLPs in host cells by in situ immunogold detection, electron microscopy and immunofluorescence. Characteristics of these EV71 VLPs were studied using a variety of immunological and physicochemical techniques, which aimed to reveal that the purified EV71 VLPs have good morphology and structure consistent with natural EV71 empty capsids. Results of the amino acid analysis, SDS-PAGE, Western blotting and high-performance liquid chromatography confirmed the high purity of the EV71 VLPs. However the sedimentation coefficient of the VLPs showed that they were smaller than that of secreted EV71 VLPs purified by discontinuous cesium chloride density gradients, they were similar to the empty capsids of natural EV71 virions reported previously. Combined with the previous study that EV71 VLPs purified by a multistep chromatography process were able to elicit strong humoral immune responses in mice, our results further supported the conclusion that our EV71 VLPs had well-preserved molecular and structural characteristics. The EV71 VLPs produced from the baculovirus expression system and purified by a multistep chromatography process displayed key structural and immunological features, which would contribute to their efficacy as a HFMD vaccine.


Porphyrin Grafting on a Mercapto-Equipped Zr(IV)-Carboxylate Framework Enhances Photocatalytic Hydrogen Production.
We employ facile aromatic nucleophilic substitution between the mercapto (-SH) and arylfluoro (Ar-F) groups to achieve extensive and robust cross-linking of a coordination host by porphyrin guests that also serve the purpose of versatile postsynthetic functionalization. For this, a tritopic linker with three trident-like thiol-flanked carboxyl units are reacted with ZrOCl2·8H2O to afford a two-dimensional (3,6-connected) net. The wide aperture of the porous framework solid, together with its stability in both air and boiling water, facilitates the entry of bulky metalloporphyrin guests and the subsequent property studies. On the porphyrin side, four pentafluorophenyl (C6F5-) groups offer multiple fluoro groups to facilitate their replacement by the thiol groups from the host net. The inserted metalloporphyrin bridges impart to the metal-organic framework (MOF) host stable and recyclable activities for photocatalytic hydrogen production. We also disclose an improvement in synthetic methodology, in which BBr3 is used to simultaneously cleave the ester and benzyl thioether groups to more efficiently access thiol-equipped carboxylic acid building block.


Electrostatic effects on the kinetics of bound enzymes.
1. The effect of the interaction between the charged matrix and substrate on the kinetic behaviour of bound enzymes was investigated theoretically. 2. Simple expression is derived for the apparent Km. 3. The apparent Km can only be used for the characterization of the electrostatic effect of the ionic strength does not vary with the substrate concentration. 4. The deviations from Michaelis-Menton kinetics are graphically illustrated for cases when the ionic strength varies with the substrate concentration. 5. The inhibition of the bound enzyme by a charged inhibitor at constant ionic strength is characterized by an apparent Ki. 6. When both the inhibitor concentration and the ionic strength change there is no apparent Ki, and the inhibition profile is graphically illustrated for this case. 7. Under certain conditions the electrostatic effects manifest thenselves in a sigmoidal dependence of the enzyme activity on the concentration of the substrate or inhibitor.


Experimental and Monte Carlo study of the effect of the presence of dry air, cortical bone inhomogeneities and source position on dose distribution of the mHDR-v2 source.
Recently it was data wise established that there is a considerable dose difference due to source position from the surface of the patient, and due to the presence of inhomogeneities. It aims at to find out the dose difference due to source position, and inhomogenieties in water phantom of high dose rate (HDR)(192) Ir mHDR-v2 source by experiment and by Monte Carlo (MC) simulation GEANT4 code. The measured study of the source was done using an in-air ionization chamber, water phantom while the calculated study was done by modeling the water phantom and its water, inhomogeneities, position of source, and points of calculation. The measured and calculated dose differences are 5.48 to 6.46% and 5.43 to 6.44% respectively higher in the presence of dry air and 4.40 to 4.90% and 4.38 to 4.88% respectively lower in the presence of cortical bone. However, for the study of the effect of source position on dose distribution, when the source was positioned at a 1 cm distance from the surface of water phantom, the near points between 1 cm and 2 cm are 2 to 3.5% and 2.1-3.7% underdose and for distant points from 3 cm to 8 cm from the source are 4 to 15% and 4.1 to 15.8% underdose for measured and calculated studies, respectively, to the dose when the source was positioned at midpoint of water phantom. These results can be used in the treatment planning system.


Isorhynchophylline attenuates proliferation and migration of synovial fibroblasts via the FOXC1/β-catenin axis.
Rheumatoid arthritis (RA) is a common type of chronic inflammatory disease. Elucidating the mechanism of fibroblast-like synovial (FLS) as a pathologic factor in RA may address the urgent medical requirement for the treatment of RA. Isorhynchophylline (IRN) is a tetracyclic hydroxyindole alkaloid isolated from uncinaria, which has multiple biological activities and affects the progression of osteoarthritis. However, the role of IRN in rheumatoid arthritis remains unclear. Herein, our study aimed to elucidate the potential effect of IRN on RA and reveal its mechanism. Human FLS cell line MH7A cells were stimulated with TNF-α for 24 h to construct a cell model. CCK-8, Edu, wound healing, as well as transwell assays were conducted to detect the effects of IRN on cell proliferation and motility. ELISA and Immunoblot assays were further performed to detect the production of pro-inflammatory factors and the expression levels of MMPs. Immunoblot and Immunostaining assays were conducted to uncover the mechanism. ELISA, H&E staining, and Immunoblot assays were used to confirm the effects of IRN on RA in a CIA rat model. We revealed that IRN restrained TNF-α-stimulated MH7A cell proliferation and motility. In addition, IRN blocked the production of pro-inflammatory factors and MMPs in TNF-α-stimulated-MH7A cells. We further found that IRN restrained FOXC1/β-catenin axis, and improved MH7A cell proliferation as well as migration via the FOXC1/β-catenin axis. IRN restores CIA by inhibiting pro-inflammatory cytokines in synovial tissues. In summary, IRN attenuates proliferation and migration of FLS in RA via the FOXC1 mediated β-catenin axis.


Using the 6-minute walk test to predict disability-free survival after major surgery.
The 6-min walk test (6MWT) is a common means of functional assessment. Its relationship to disability-free survival (DFS) is uncertain. This sub-study of the Measurement of Exercise Tolerance for Surgery study had co-primary outcome measures: correlation of the preoperative 6MWT distance with 30 day quality of recovery (15-item quality of recovery) and 12 month WHO Disability Assessment Schedule scores. The prognostic utility of the 6MWT and other risk assessment tools for 12 month DFS was assessed with logistic regression and receiver-operating-characteristic-curve analysis. Of 574 patients recruited, 567 (99%) completed the 6MWT. Twelve months after surgery, 16 (2.9%) patients had died and 444 (77%) had DFS. The 6MWT correlated weakly with 30 day 15-item quality of recovery (ρ=0.14; P=0.001) and 12 month WHO Disability Assessment Schedule (ρ=-0.23; P<0.0005) scores. When the cohort was split into 6MWT distance tertiles, the adjusted odds ratio of low vs high tertiles for DFS was 3.13 [95% confidence interval (CI): 1.54-6.35]. The only independent variable predictive of DFS was the Duke Activity Status Index (DASI) score (adjusted odds ratio: 1.06; P<0.0005). The area under the receiver-operating-characteristic curve for DFS was 0.63 (95% CI: 0.57-0.70) for the 6MWT, 0.60 (95% CI: 0.53-0.67) for cardiopulmonary-exercise-testing-derived peak oxygen consumption, and 0.70 (95% CI: 0.64-0.76) for the DASI score. Of the risk assessment tools analysed, the DASI was the most predictive of DFS. The 6MWT was safe and comparable with cardiopulmonary exercise testing for all predictive assessments. Future research should aim to determine the optimal 6MWT distance thresholds for risk prediction.


Local Drug Infiltration Analgesia During Knee Surgery to Reduce Postoperative Pain in Rats.
There is increasing interest in local infiltration analgesia (LIA) to reduce postoperative pain with knee surgery. Despite widespread use of LIA, wide variations in drug combinations, infiltration techniques, and the concomitant use of systemic analgesics have made it difficult to determine the optimal drug combination for LIA.Using a previously validated animal knee surgery model, we aimed to determine the optimal combination of medications to reduce postoperative pain, and the best anatomical location and timing for local drug injection during surgery. Knee surgery was performed in an adult rat model under isoflurane anesthesia. During surgery, combinations of bupivacaine, ketorolac, dexamethasone, and morphine were injected around the knee and compared to saline placebo. Similar medications were injected systemically as a comparator group. Postoperative pain was assessed by measuring spontaneous rearing activity. Injections were given after bone drilling and/or just before wound closure. The 3-drug LIA combination of bupivacaine, ketorolac, and dexamethasone increased rearing (decreased pain) at 2 hours (P = 0.0198) and 24 hours (P = 0.0384) postsurgery compared to saline. The same drugs injected systemically had no effect. The ketorolac/dexamethasone combination for LIA was also effective at 2 hours (P = 0.0006) and 24 hours (P = 0.0279), and ketorolac alone reduced pain at 2 hours (P = 0.0045). Bupivacaine alone was less effective, and the addition of morphine had no effect. The 3-drug combination infiltrated just after creating holes in bone was more effective than when given into the wound just before wound closure. Our animal study suggests that clinical trials with LIA combinations of local anesthetic, nonsteroidal anti-inflammatory drug, and corticosteroid might be useful for reducing postoperative pain after knee surgery, with the nonsteroidal anti-inflammatory drug having the greatest effect.Perioperative physicians should consider delivering LIA earlier during the procedure as opposed to solely at the time of wound closure.


Unsupervised Person Re-Identification by Deep Asymmetric Metric Embedding.
Person re-identification (Re-ID) aims to match identities across non-overlapping camera views. Researchers have proposed many supervised Re-ID models which require quantities of cross-view pairwise labelled data. This limits their scalabilities to many applications where a large amount of data from multiple disjoint camera views is available but unlabelled. Although some unsupervised Re-ID models have been proposed to address the scalability problem, they often suffer from the view-specific bias problem which is caused by dramatic variances across different camera views, e.g., different illumination, viewpoints and occlusion. The dramatic variances induce specific feature distortions in different camera views, which can be very disturbing in finding cross-view discriminative information for Re-ID in the unsupervised scenarios, since no label information is available to help alleviate the bias. We propose to explicitly address this problem by learning an unsupervised asymmetric distance metric based on cross-view clustering. The asymmetric distance metric allows specific feature transformations for each camera view to tackle the specific feature distortions. We then design a novel unsupervised loss function to embed the asymmetric metric into a deep neural network, and therefore develop a novel unsupervised deep framework named the DEep Clustering-based Asymmetric MEtric Learning (DECAMEL). In such a way, DECAMEL jointly learns the feature representation and the unsupervised asymmetric metric. DECAMEL learns a compact cross-view cluster structure of Re-ID data, and thus help alleviate the view-specific bias and facilitate mining the potential cross-view discriminative information for unsupervised Re-ID. Extensive experiments on seven benchmark datasets whose sizes span several orders show the effectiveness of our framework.


Preoperative characteristics of patients with presumed pancreatic cancer but ultimately benign disease: a multicenter series of 344 pancreatoduodenectomies.
Preoperative differentiation between malignant and benign pancreatic tumors can be difficult. Consequently, a proportion of patients undergoing pancreatoduodenectomy for suspected malignancy will ultimately have benign disease. The aim of this study was to compare preoperative clinical and diagnostic characteristics of patients with unexpected benign disease after pancreatoduodenectomy with those of patients with confirmed (pre)malignant disease. We performed a multicenter retrospective cohort study in 1,629 consecutive patients undergoing pancreatoduodenectomy for suspected malignancy between 2003 and 2010 in 11 Dutch centers. Preoperative characteristics were compared in a benign:malignant ratio of 1:3. Malignant cases were selected from the entire cohort by using a random number list. A multivariable logistic regression prediction model was constructed to predict benign disease. Of 107 patients (6.6 %) with unexpected benign disease after pancreatoduodenectomy, 86 fulfilled the inclusion criteria and were compared with 258 patients with (pre)malignant disease. Patients with benign disease presented more often with pain (56 vs. 38 %; P = 0.004), but less frequently with jaundice (60 vs. 80 %; P < 0.01), a pancreatic mass (13 vs. 54 %, P < 0.001), or a double duct sign on computed tomography (21 vs. 47 %; P < 0.001). In a prediction model using these parameters, only 19 % of patients with benign disease were correctly predicted, and 1.4 % of patients with malignant disease were missed. Nearly 7 % of patients undergoing pancreatoduodenectomy for suspected malignancy were ultimately diagnosed with benign disease. Although some preoperative clinical and imaging characteristics might indicate absence of malignancy, their discriminatory value is insufficient for clinical use.


Sorting colloidal particles into multiple channels with optical forces: prismatic optical fractionation.
Brownian particles drifting through a periodically structured force landscape can become entrained by the landscape's symmetries. What direction a particular particle takes can depend strongly on subtle variations in its physical properties. Consequently, a homogeneously structured force field can sort a mixture of particles into spatially separated fractions, much as an optical prism refracts light into its component wavelengths. When the force landscape is implemented with structured light fields, such continuous multichannel sorting may be termed prismatic optical fractionation. We describe experimental and numerical studies of colloidal spheres' transport through periodic arrays of optical tweezers, which reveal an important role for three-dimensional motion in determining a drifting particle's fate. These studies also demonstrate sorting on the basis of statistically locked-in transport, in which brownian fluctuations contribute to direction selection.


Periodontal Treatment and the Risks of Cardiovascular Disease in Patients with Type 2 Diabetes: A Retrospective Cohort Study.
Objective Periodontal disease may predispose individuals to cardiovascular disease (CVD). Diabetes mellitus, especially in patients with severe periodontitis, increases the risk of CVD mortality. However, the outcomes of periodontal therapy vary among the different treatment modalities. We aim to investigate whether periodontal treatment could influence the occurrence of CVD in patients with type 2 diabetes and periodontal problems. Methods A retrospective cohort study was conducted based on a dataset released by Taiwan National Health Insurance (NHI). The dataset was composed of randomly sampled, newly diagnosed diabetic patients who received insurance benefits from 1999 to 2001; patients who were younger than 18 years of age or who already had CVD before 1999 were excluded. The NHI code was used to identify the treatments, including subgingival curettage and flap operations. The patients' demographic variables were matched using a 1:4 propensity score. All of the subjects were followed up until the onset of CVD, or December 31, 2011. A Cox proportional hazards regression analysis was performed to evaluate the effects of periodontal treatment on the rates of myocardial infarction, heart failure and stroke. Results Three thousand thirty-nine and 12,156 diabetic subjects were classified into the advanced periodontal treatment group and the non-advanced periodontal treatment group, respectively. The Cox proportional hazards analysis revealed that although the overall incidence of CVD was not significantly improved (Hazard ratio, HR 0.95; 95% CI 0.90-1.01), advanced periodontal treatment reduced the rates of myocardial infarction (HR 0.92; 95% CI 0.85-0.99) and heart failure (HR 0.60; 95% CI 0.45-0.80). There was no significance difference in the incidence of stroke (HR 0.95; 95% CI 0.85-1.06). Conclusion Advanced periodontal therapy lowers the rate of CVD, especially myocardial infarction and heart failure. Dental management has a beneficial effect on the health of patients with type 2 diabetes.


Ventral lateral and DN1 clock neurons mediate distinct properties of male sex drive rhythm in Drosophila.
Male sex drive rhythm (MSDR) in Drosophila is a circadian behavior only observed in the social context of male-female pairs. In the presence of a female, males exhibit long periods of courtship activity with a pronounced rest phase at dusk, although isolated males exhibit an activity peak at dusk. The molecular mechanisms regulating the switch between these activity patterns are unknown. Here, we genetically manipulate the molecular clock in different subsets of neurons and find that proper oscillation of the molecular clock in ventral lateral neurons is essential for MSDR. These neurons express pigment-dispersing factor, the lack of which disrupts MSDR. Furthermore, we show that a cluster of dorsal neurons (DN1s) requires the molecular clock to synchronize the trough phase at dusk in MSDR and to establish the evening peak in single fly locomotor rhythm (SLR). Finally, we provide evidence that DN1s exert their roles in MSDR and SLR via distinct signaling pathways.


Faculty Decision Making in Ad Hoc Entrustment of Pediatric Critical Care Fellows: A National Case-Based Survey.
Phenomenon: Ad hoc entrustment decisions reflect a clinical supervisor's estimation of the amount of supervision a trainee needs to successfully complete a task in the moment. These decisions have important consequences for patient safety, trainee learning, and preparation for independent practice. Determinants of these decisions have previously been described but have not been well described for acute care contexts such as critical care and emergency medicine. The ad hoc entrustment of trainees caring for vulnerable patient populations is a high-stakes decision that may differ from other contexts. Critically ill patients and children are vulnerable patient populations, making the ad hoc entrustment of a pediatric critical care medicine (PCCM) fellow a particularly high-stakes decision. This study sought to characterize how ad hoc entrustment decisions are made for PCCM fellows through faculty ratings of vignettes. The authors investigated how acuity, relationship, training level, and task interact to influence ad hoc entrustment decisions. Approach: A survey containing 16 vignettes that varied by four traits (acuity, relationship, training level, and task) was distributed to U.S. faculty of pediatric critical care fellowships in 2020. Respondents determined an entrustment level for each case and provided demographic data. Entrustment ratings were dichotomized by "high entrustment" versus "low entrustment" (direct supervision or observation only). The authors used logistic regression to evaluate the individual and interactive effects of the four traits on dichotomized entrustment ratings. Findings: One hundred seventy-eight respondents from 30 institutions completed the survey (44% institutional response rate). Acuity, relationship, and task all significantly influenced the entrustment level selected but did not interact. Faculty most frequently selected "direct supervision" as the entrustment level for vignettes, including for 24% of vignettes describing fellows in their final year of training. Faculty rated the majority of vignettes (61%) as "low entrustment." There was no relationship between faculty or institutional demographics and the entrustment level selected. Insights: As has been found in summative entrustment for pediatrics, internal medicine, and surgery trainees, PCCM fellows often rated at or below the "direct supervision" level of ad hoc entrustment. This may relate to declining opportunities to practice procedures, a culture of low trust propensity among the specialty, and/or variation in interpretation of entrustment scales.


Kinetics of GABAB autoreceptor-mediated suppression of GABA release in rat insular cortex.
Release of GABA is controlled by presynaptic GABA receptor type B (GABA(B)) autoreceptors at GABAergic terminals. However, there is no direct evidence that GABA(B) autoreceptors are activated by GABA release from their own terminals, and precise profiles of GABA(B) autoreceptor-mediated suppression of GABA release remain unknown. To explore these issues, we performed multiple whole-cell, patch-clamp recordings from layer V rat insular cortex. Both unitary inhibitory and excitatory postsynaptic currents (uIPSCs and uEPSCs, respectively) were recorded by applying a five-train depolarizing pulse injection at 20 Hz. In connections from both fast-spiking (FS) and non-FS interneurons to pyramidal cells, the GABA(B) receptor antagonist CGP 52432 had little effect on the initial uIPSC amplitude. However, uIPSCs, responding to later pulses, were effectively facilitated. This CGP 52432-induced facilitation was prominent in the fourth uIPSCs, which were evoked 150 ms after the first uIPSC. The facilitation of uIPSCs was accompanied by an increase in the paired-pulse ratio. In addition, analysis of the coefficient of variation suggests the involvement of presynaptic mechanisms in CGP 52432-induced uIPSC facilitation. Paired-pulse stimulation (interstimulus interval = 150 ms) of presynaptic FS cells revealed that the second uIPSC was also facilitated by CGP 52432, which had little effect on the amplitude and interevent interval of miniature IPSCs. In contrast, uEPSCs, responding to all five stimulations of a presynaptic pyramidal cell, were less affected by CGP 52432. These results suggest that a single presynaptic action potential is sufficient to activate GABA(B) autoreceptors and to suppress GABA release in the cerebral cortex.


A map of the phosphoproteomic alterations that occur after a bout of maximal-intensity contractions.
Mechanical signals play a critical role in the regulation of muscle mass, but the molecules that sense mechanical signals and convert this stimulus into the biochemical events that regulate muscle mass remain ill-defined. Here we report a mass spectrometry-based workflow to study the changes in protein phosphorylation that occur in mouse skeletal muscle 1 h after a bout of electrically evoked maximal-intensity contractions (MICs). Our dataset provides the first comprehensive map of the MIC-regulated phosphoproteome. Using unbiased bioinformatics approaches, we demonstrate that our dataset leads to the identification of many well-known MIC-regulated signalling pathways, as well as to a plethora of novel MIC-regulated events. We expect that our dataset will serve as a fundamentally important resource for muscle biologists, and help to lay the foundation for entirely new hypotheses in the field. The maintenance of skeletal muscle mass is essential for health and quality of life. It is well recognized that maximal-intensity contractions, such as those which occur during resistance exercise, promote an increase in muscle mass. Yet, the molecules that sense the mechanical information and convert it into the signalling events (e.g. phosphorylation) that drive the increase in muscle mass remain undefined. Here we describe a phosphoproteomics workflow to examine the effects of electrically evoked maximal-intensity contractions (MICs) on protein phosphorylation in mouse skeletal muscle. While a preliminary phosphoproteomics experiment successfully identified a number of MIC-regulated phosphorylation events, a large proportion of these identifications were present on highly abundant myofibrillar proteins. We subsequently incorporated a centrifugation-based fractionation step to deplete the highly abundant myofibrillar proteins and performed a second phosphoproteomics experiment. In total, we identified 5983 unique phosphorylation sites of which 663 were found to be regulated by MIC. GO term enrichment, phosphorylation motif analyses, and kinase-substrate predictions indicated that the MIC-regulated phosphorylation sites were chiefly modified by mTOR, as well as multiple isoforms of the MAPKs and CAMKs. Moreover, a high proportion of the regulated phosphorylation sites were found on proteins that are associated with the Z-disc, with over 74% of the Z-disc proteins experiencing robust changes in phosphorylation. Finally, our analyses revealed that the phosphorylation state of two Z-disc kinases (striated muscle-specific serine/threonine protein kinase and obscurin) was dramatically altered by MIC, and we propose ways these kinases could play a fundamental role in skeletal muscle mechanotransduction.


Age-independent rise of inflammatory scores may contribute to accelerated aging in multi-morbidity.
Aging is associated with an increase in a chronic, low-grade inflammation. This phenomenon, termed "inflammaging" is also a risk factor for both morbidity and mortality in the elderly. Frequent co-occurrence of chronic diseases, known as multi-morbidity, may be explained by interconnected pathophysiology of these conditions, most of which depend on its inflammatory component. Here we present an analysis of the U.S. National Health and Nutrition Examination Survey data collected between 1999 and 2008, for the presence, and the number, of chronic diseases along with HDL-cholesterol, C-reactive protein, white blood cell count, lymphocyte percent, monocyte percent, segmented neutrophils percent, eosinophils percent, basophils percent, and glycohemoglobin levels. Importantly, even after adjustment for age and BMI, many inflammatory markers continued to be associated to multi-morbidity. C-reactive protein (CRP) levels and Glasgow Prognostic Score (GPS) were most dramatically increased in parallel with an accumulation of chronic diseases, and may be utilized as multi-morbidity predictors. These observations point at background inflammation as direct, age-independent contributor to an accumulation of the disease burden. Our findings also suggest a possibility that systemic inflammation associated with chronic diseases may explain accelerated aging phenomenon previously observed among the patients with heavy disease burden.


Determinants of soft budget constraints: How public debt affects hospital performance in Austria.
Soft budget constraints (SBCs) undermine reforms to increase hospital service efficiency when hospital management can count on being bailed out by (subnational) governments in case of deficits. Using cost accounting data on publicly financed, non-profit hospitals in Austria from 2002 to 2015, we analyse the association between SBCs and hospital efficiency change in a setting with negligible risk of hospital closure in a two-stage study design based on bias-corrected non-radial input-oriented data envelopment analysis and ordinary least squares regression. We find that the European debt crisis altered the pattern of hospital efficiency development: after the economic crisis, hospitals in low-debt states had a 1.1 percentage point lower annual efficiency change compared to hospitals in high-debt states. No such systematic difference is found before the economic crisis. The results suggest that sudden exogenous shocks to public finances can increase the budgetary pressure on publicly financed institutions, thereby counteracting a pre-existing SBC.


Thymoquinone reduces migration and invasion of human glioblastoma cells associated with FAK, MMP-2 and MMP-9 down-regulation.
Glioblastoma represent the most frequent primary tumors of the central nervous system and remain among the most aggressive human cancers as available therapeutic approaches still fail to contain their invasiveness. Many studies have reported elevated expression of the Focal Adhesion Kinase (FAK) protein in glioblastoma, associated with an increase in the rates of both migration and invasion. This designates FAK as a promising target to limit invasiveness in glioblastoma. Thymoquinone (TQ), the main phytoactive compound of Nigella sativa has shown remarkable anti-neoplasic activities on a variety of cancer cells. Here, we studied the anti-invasive and anti-migratory effects of TQ on human glioblastoma cells. The results obtained indicated that TQ treatment reduced migration, adhesion and invasion of both U-87 and CCF-STTG1 cells. This was accompanied by a drastic down-regulation of FAK, associated with a reduction of ERK phosphorylation as well as MMP-2 and MMP-9 secretion. This study provides new data on FAK regulation by a natural product (TQ) which could be of a great value for the development of novel therapies in glioblastoma.


Prehospital delay in patients with acute coronary heart disease: concordance between patient interviews and medical records.
Patient-associated delay in seeking medical care in the setting of acute coronary disease is assuming increasing importance as the benefits of reperfusion therapies become more time dependent. Given the importance of accurate information concerning prehospital delay, we examined the extent of concordance between information reported by patients in structured interviews by hospital staff nurses compared with information about time of acute symptom onset as recorded in the medical record. Data were obtained from 1137 patients with a discharge diagnosis of coronary heart disease who were admitted to six coronary care units in the Minneapolis-St. Paul metropolitan area. The average and median durations of prehospital delay were similar as reported in the structured personal interviews and through the review of medical records for the respective disease groups. The extent of individual level of agreement of delay time was considerably poorer, however. The Pearson correlation coefficients on the logarithmically transformed data were 0.48, 0.50, and 0.59 for persons with acute myocardial infarction, unstable angina, and chronic coronary disease, respectively, in comparing data noted in the medical record with that obtained in the personal interviews concerning prehospital delay time. These results suggest good agreement between personal interviews and medical record accounts in characterizing the average length of prehospital delay at the aggregate level but considerably less agreement at the individual patient level.


Central Reading of Endoscopy Endpoints in Inflammatory Bowel Disease Trials.
Central reading of endoscopy (CROE) is crucial in determining who qualifies for a trial but also has a role, independent of the selected scoring system, in decreasing measurement noise that can obscure separation between placebo and active drug. Benefits of CROE may not be independent of the method chosen, and controversy exists about the ideal approach. Literature review and concept development. Components to be considered in the reading algorithm are blinding, number of central readers, independent voting versus consensus panel, video recordings versus still images, and involvement of the site reader. Key concepts considered are endpoints, bias, power, and sample size derived from the Food and Drug Administration and European Medicines Agency guidelines, as well as the technological requirements and recruitment, qualification, and revalidation of central readers as applied to CROE. Recording and CROE should be standardized, and an imaging charter developed with research on the different components and its overall impact.


Retention of fundamental surgical skills learned in robot-assisted surgery.
Evaluation of the learning curve for robotic surgery has shown reduced errors and decreased task completion and training times compared with regular laparoscopic surgery. However, most training evaluations of robotic surgery have only addressed short-term retention after the completion of training. Our goal was to investigate the amount of surgical skills retained after 3 months of training with the da Vinci™ Surgical System. Seven medical students without any surgical experience were recruited. Participants were trained with a 4-day training program of robotic surgical skills and underwent a series of retention tests at 1 day, 1 week, 1 month, and 3 months post-training. Data analysis included time to task completion, speed, distance traveled, and movement curvature by the instrument tip. Performance of the participants was graded using the modified Objective Structured Assessment of Technical Skills (OSATS) for robotic surgery. Participants filled out a survey after each training session by answering a set of questions. Time to task completion and the movement curvature was decreased from pre- to post-training and the performance was retained at all the corresponding retention periods: 1 day, 1 week, 1 month, and 3 months. The modified OSATS showed improvement from pre-test to post-test and this improvement was maintained during all the retention periods. Participants increased in self-confidence and mastery in performing robotic surgical tasks after training. Our novel comprehensive training program improved robot-assisted surgical performance and learning. All trainees retained their fundamental surgical skills for 3 months after receiving the training program.


[Olfactory groove meningiomas. Radical microsurgical treatment through the bifrontal approach].
To describe the microsurgical technique for the radical removal of olfactory groove meningiomas through the bifrontal approach. To review the diagnostic elements to be taken into account in the selection of the surgical approach to these tumours. A microsurgical series of 35 olfactory groove meningiomas operated on through a bifrontal craniotomy is reviewed. The mean tumoral volume was 85cc (4.4cm diameter). A relevant peritumoral brain edema was found in 65.7% of cases, hyperostosis in the implantation base in 80% and paranasal sinus invasion in 28.6%. A Sipmson grade 1 resection was achieved in every case. A patient died due to a postoperative pneumonia. Postoperative hospitalization time was between 3 and 20 days and at discharge all patients had a Glasgow Outcome Scale grade 4-5. The mean follow-up was 55.2 months. Two patients had postoperative transient rhinolicuorrhea and an additional patient developed hydrocephalus. An asymptomatic recurrence have been identified in a patient four years after surgery. In our experience the bifrontal approach allowed the radical removal of huge olfactory groove meningiomas. The microdissection of the anterior cerebral artery A2 segments is possible thanks to the arachnoidal plane between vessels and tumor. Tumoral blood flow is secured by the early approaching of the base of the tumor and preoperative embolization is not necessary. Bifrontal approach allows an aggressive treatment of the hyperostosis, bone infiltration and paranasal sinus invasion. Anterior fossa reconstruction is done using a vascularized periosteal flap.


Nail lichen planus treatment safety.
Topical therapies for nail lichen planus (clobetasol propionate, topical tacrolimus, bath-PUVA), intralesional treatment (triamcinolone), and systemic treatment (corticosteroids, retinoids, small molecule inhibitors (jak/stat inhibitors)), TNF-alpha inhibitors (etanercept), systemic immunomodulators (oral calcineurin inhibitors, mycophenolate mophetil), and antimalarials (chloroquine), each with unique safety profiles and considerations. Herein, we discuss common and uncommon adverse events, as well as utilization for special populations, including pregnant and pediatric patients.


Cancer in the electric power industry.
A cohort study of 3358 power linesmen and 6703 power station operators classified in the 1960 population census in Sweden was undertaken. The cancer incidence between 1961 and 1979 was calculated in relation to all male blue collar workers. The relative risk of cancer (all sites) was 1.1, (1.0-1.2, 90% confidence limit) in the group of power linesmen and 1.0 (0.9-1.0) for power station operators. No consistent excess risk of leukaemia or brain tumour was found in the two occupational groups. In both groups a tendency towards an excess risk of cancer of the urinary organs was found.


Pulsed arterial spin labelling at ultra-high field with a B 1 (+) -optimised adiabatic labelling pulse.
Arterial spin labelling (ASL) techniques benefit from the increased signal-to-noise ratio and the longer T 1 relaxation times available at ultra-high field. Previous pulsed ASL studies at 7 T concentrated on the superior regions of the brain because of the larger transmit radiofrequency inhomogeneity experienced at ultra-high field that hinders an adequate inversion of the blood bolus when labelling in the neck. Recently, researchers have proposed to overcome this problem with either the use of dielectric pads, through dedicated transmit labelling coils, or special adiabatic inversion pulses. We investigate the performance of an optimised time-resampled frequency-offset corrected inversion (TR-FOCI) pulse designed to cause inversion at much lower peak B 1 (+) . In combination with a PICORE labelling, the perfusion signal obtained with this pulse is compared against that obtained with a FOCI pulse, with and without dielectric pads. Mean grey matter perfusion with the TR-FOCI was 52.5 ± 10.3 mL/100 g/min, being significantly higher than the 34.6 ± 2.6 mL/100 g/min obtained with the FOCI pulse. No significant effect of the dielectric pads was observed. The usage of the B 1 (+) -optimised TR-FOCI pulse results in a significantly higher perfusion signal. PICORE-ASL is feasible at ultra-high field with no changes to operating conditions.


Fast liquid chromatography-tandem mass spectrometry method for routine assessment of irinotecan metabolic phenotype.
Irinotecan (CPT-11) is an anticancer drug with a complex in vivo metabolism widely used in the treatment of colon cancer. The assessment of the CPT-11 metabolic phenotype is an important clinical component for personalizing the administered dose. In this study, we report the development of a rapid and sensitive liquid chromatography-tandem mass spectroscopy method for the simultaneous quantitation of CPT-11 and its major metabolites generated by hydrolysis (SN-38, active metabolite) or through oxidative (NPC and APC) and glucuronidation (SN-38G) pathways. The method used a small volume of plasma and is based on simple protein plasma precipitation with two volumes of acetonitrile containing camptothecine as an internal standard. Analytes extracted in the supernatant fraction were converted to the lactone form and further separated by an acetonitrile gradient on a Kinetex C18 (50 × 2 mm) column in the presence of 0.01% formic acid. The quantitative measurement was performed by an API 4000/Qtrap operating in the triple-quadruple mode. The Multiple Reaction Monitoring transitions were: m/z 587→167 for CPT-11, m/z 393→349 for SN-38, m/z 619→393 for APC, m/z 519→393 for NPC, and m/z 569→393 for SN-38G. The mean overall recovery in plasma was in the range of 78% to 86% with a low matrix suppression effect (9.0% or less). The lower limit of quantitation was 0.2 ng/mL for NPC and SN-38 and 0.5 ng/mL for SN-38G, APC, and CPT-11. Linearity was checked up to 2000 ng/mL, whereas the intra- and interday accuracy and precision for both the parent drug and its metabolites was -3.7% to 13.8% and 3.7% to 13.1%, respectively. The method proved to be robust and suitable for therapeutic drug monitoring as well as for pharmacogenetic-pharmacokinetic correlation studies.


Relevance of the Pediatric Powered Wheelchair Screening Test for children with cerebral palsy.
In a previous study we developed a cognitive assessment battery called the Pediatric Powered Wheelchair Screening Test (PPWST) to help clinicians determine a young child's readiness to drive a powered wheelchair. The current multicenter study sought to determine: (1) whether the PPWST is appropriate for use in a population of children with cerebral palsy (CP) who use joysticks to drive their wheelchair; (2) whether two additional factors (symbolic representation and coping) would increase the predictive power of the PPWST for this group and for children with orthopedic or neuromuscular disabilities only; and (3) whether the test was appropriate for children with severe motor impairments who use switches to control their wheelchair. Fifty children (27 males, 23 females) between the ages of 21 months and 6 years 11 months participated. Twenty-six children (mean age 4 years 4 months, SD 15 months) had triplegic or tetraplegic CP and 24 children (mean age 27 months, SD 5 months) had orthopedic or neuromuscular disabilities. Sixty-nine per cent of children had some limited form of mobility (such as rolling or scooting) but none was a functional ambulator. Each child was assessed with the PPWST and with measures of symbolic representation and coping. After six wheelchair training sessions, driving ability was scored. The PPWST was found to be predictive of functional driving ability for children with CP who used a joystick to control their wheelchair. Assessment of symbolic representation skills increased the predictive power for this group but not for children with orthopedic or neuromuscular disabilities; coping scores did not increase the predictive power for either group. The PPWST accounted for only 20% of the variance in overall driving skills for switch users, and thus is not yet considered an adequate screening device for this group. The PPWST is designed to help clinicians determine whether a child currently has the specific cognitive skills found to be related to powered wheelchair driving but is not intended to be used exclusively to determine whether or not a child is ultimately a candidate for powered mobility.


Visual impairment and amblyopia in Malaysian pre-school children - The SEGPAEDS study.
Little is known regarding the extent of visual impairment amongst pre-school children in Malaysia. To determine the prevalence of visual impairment and amblyopia in Malaysian preschool children. A cross-sectional, population-based study was conducted on children aged four to six years from 51 participating kindergartens in the district of Segamat, Johor, Malaysia from 20 March 2016 to 6 April 2016. All subjects had initial eye screening consisting of LogMar visual acuity, orthoptics examination and Spot vision screener assessment. Subjects who failed the initial eye screening were invited for a formal eye assessment consisting of cycloplegic refraction and a comprehensive ocular examination. Definitions of visual impairment and amblyopia were based on the Multi-Ethnic Pediatric Eye Disease Study criteria. A total of 1287 children were recruited. Mean subject age was 5.03 (SD:0.77) and males represented 52.3% of subjects. Subjects by ethnicity were Malay (54.8%), Chinese (27.7%), Indian (15.6%) and Orang Asli (1.9%). Formal eye assessment was required for 221 subjects and 88.8% required ophthalmic intervention. Refractive error, representing 95.4% of diagnosed ocular disorders, comprised of astigmatism (84%), myopia (9%) and hypermetropia (6.9%). With-the-rule astigmatism was present in 93.4% of the subjects with astigmatism. Visual impairment was present in 12.5% of our subjects, with 61% having bilateral visual impairment. Of the subjects with visual impairment, 59.1% had moderate visual impairment. The prevalence of amblyopia was 7.53%, and 66% of the amblyopic subjects had bilateral amblyopia. Our study highlights an urgent need for initiation of preschool vision screening in Malaysia.


Transferrin and the transferrin receptor: of magic bullets and other concerns.
Transferrin (Trf) is a highly conserved serum glycoprotein mostly known for its iron transport capacity. As iron is an indispensable nutrient for cell division, Trf and its receptor have long been used as targets of pharmacological intervention mostly for cancer therapy and for diagnosis in inflammation. In recent years several independent pieces of work including data from our group, indicated that Trf can also have an iron independent role in the immune system. In this article new emerging roles of Trf and its receptor on iron independent processes and in drug delivery are reviewed.


Identification of morphine accumulation in the rat embryo central nervous system: a c14-morphine administration study.
Previous studies have shown that morphine consumption during pregnancy may cause delay or defect of embryo development or abnormal nervous system function in the human and animal models. In the present study, the highest density of morphine accumulation in the central nervous system of rat embryos was evaluated using C14-morphine. Female Wistar rats (W 170-200 g) used and were crossed with male rats and coupling time was recorded (Embryonic day 0-E0). Experimental groups received 0.05 mg/ml of C14-morphine in drinking water daily. On the 10(th) and 17(th) days of pregnancy, pregnant rats were anesthetized and the embryos with these uterus and placenta were surgically removed and were fixed in formalin 10% for 4 week. Then the embryos were processed, sectioned in 25 μm and 5 μm thicknesses, fixed on the glasses for further evaluations. The sectioned in 25, the glasses were fixed on the Blanc black and white film for 6 h. Then, the films were appeared and their negatives were prepared. The sectioned in five staining hematoxylin and eosin by light microscope and MOTIC software. Our results indicated that the highest C14-morphine accumulation was observed in the vesicles and the ventricular choroid plexus (CP) of (E17) embryos, whereas, in the (E10) embryos. Highest concentration was observed in the brain vesicles and the ventricular CP. In addition, this study showed the surface area of lateral, 3(rd) and 4(th) ventricular CP in the experimental groups were increased in compared to control groups. Our results indicated that effects of morphine on reduction of embryos brain development may be due to the highest accumulation of C14-morphine in the CP and brain vesicles.


Novel Calcium-Binding Motif Stabilizes and Increases the Activity of Aspergillus fumigatus Ecto-NADase.
Nicotinamide adenine dinucleotide (NAD) is an essential molecule in all kingdoms of life, mediating energy metabolism and cellular signaling. Recently, a new class of highly active fungal surface NADases was discovered. The enzyme from the opportunistic human pathogen Aspergillus fumigatus was thoroughly characterized. It harbors a catalytic domain that resembles that of the tuberculosis necrotizing toxin from Mycobacterium tuberculosis, which efficiently cleaves NAD+ to nicotinamide and ADP-ribose, thereby depleting the dinucleotide pool. Of note, the A. fumigatus NADase has an additional Ca2+-binding motif at the C-terminus of the protein. Despite the presence of NADases in several fungal divisions, the Ca2+-binding motif is uniquely found in the Eurotiales order, which contains species that have immense health and economic impacts on humans. To identify the potential roles of the metal ion-binding site in catalysis or protein stability, we generated and characterized A. fumigatus NADase variants lacking the ability to bind calcium. X-ray crystallographic analyses revealed that the mutation causes a drastic and dynamic structural rearrangement of the homodimer, resulting in decreased thermal stability. Even though the calcium-binding site is at a long distance from the catalytic center, the structural reorganization upon the loss of calcium binding allosterically alters the active site, thereby negatively affecting NAD-glycohydrolase activity. Together, these findings reveal that this unique calcium-binding site affects the protein fold, stabilizing the dimeric structure, but also mediates long-range effects resulting in an increased catalytic rate.


Heuristic Patterns of Ethical Decision Making.
This article describes the context of ethical decision making in research and suggests that direct attention to the ways in which decisions are actually made in such environments is needed. A decision-making model based on the literature on heuristic processing is proposed and is followed by a review of the method, data, and results of the authors' research on this model. The implications of the research are developed, and a research agenda is outlined. Key findings were that competent actors do indeed process ethics problems heuristically and in ways that interweave intuitive, affective, and more rational phases. This processing does not typically follow a simple progressive pattern but evidences a sort of trial-and-error processing that is consistent with the logic of heuristic processing more generally. Finally, while diverse, participant-level processing attempts appear to follow patterns which are associated with the experience and training of the actors.